摘要
目的探讨Twist基因在结肠癌Lovo细胞奥沙利铂、氟尿嘧啶耐药中的作用。方法体外将siRNATwist、pc DNA-Twist分别转染结肠癌Lovo细胞,通过Western blot和RT-PCR方法检测细胞Twist的表达。不同浓度奥沙利铂和氟尿嘧啶、不同作用时间作用于Lovo细胞,MTT法检测细胞增殖及半数抑制浓度(IC50)。流式细胞术检测Lovo细胞凋亡情况。结果 siRNA-Twist转染Lovo细胞48 h后,细胞的Twist基因表达明显降低,而pc DNA-Twist转染后,细胞的Twist基因表达明显升高。奥沙利铂、氟尿嘧啶均呈时间、剂量依赖性抑制Lovo细胞增殖。在IC50浓度奥沙利铂和IC50浓度氟尿嘧啶作用各组Lovo细胞48 h后,siRNA-Twist组细胞生长抑制率、凋亡率明显高于空白对照组及siRNA-Control组(P<0.05),细胞对药物耐药性降低;而pc DNA-Twist组细胞生长抑制率、凋亡率明显低于空白对照组及pc DNA-Control组(P<0.05),细胞对药物耐药性升高。结论 Twist基因在结肠癌Lovo细胞奥沙利铂、氟尿嘧啶耐药中发挥正向作用。
Objective To identify the effects of Twist on the resistance of oxaliplatin and fluorouracil in colon cancer Lovo cells. Methods The expression vector of siRNA - Twist and peDNA - Twist were constructed and transfect- ed into Lovo cells by Lipofectamine. The protein and mRNA expression of Twist were detected by Western blot and RT - PCR, respectively. Methyl thiazol tetrazolium （MTF） assay was used to detect the proliferation activity of Lovo cells trea- ted by different concentrations of oxaliplatin and fluorouracil at different time points, and then decided the half maximal in- hibitory concentration （ICso）. The apoptosis rate of Lovo cells after transfection was examined by flow cytometry. Results In Lovo cells, the protein and mRNA levels of Twist were significantly reduced after transfeetion with siRNA - Twist, and which were significantly increased after transfection with peDNA - Twist. Oxaliplatin and fluorouracil inhibited the proliferation of Lovo cells both in time - and dose - dependent manners. After treated with IC50 for oxaliplatin and fluorou- racil at 48 h, the cell growth inhibition rate and apoptosis rate of siRNA - Twist group were significantly higher than blank control group and siRNA - Control group, decreasing cell resistance to drug; and which of pcDNA - Twist group were sig- nificantly higher than blank control group and pcDNA - Control group, increasing cell resistance to drug. Conclusion Twist gene plays a positive role in resistance to oxaliplatin and fluorouracil in colon cancer.
出处
《广东医学》
CAS
北大核心
2016年第22期3354-3358,共5页
Guangdong Medical Journal
基金
广东省医学科研基金资助项目(编号:B2013374)
广东省佛山市科技局立项项目(编号:201308247)
关键词
TWIST转录因子
结肠癌
耐药
Twist transcription factor
colon cancer
drug resistance
